摘要
目的观察丹皮酚对体外培养的星形胶质细胞炎性因子分泌的影响,并探讨其作用机制。方法采用神经胶质原纤维酸性蛋白(GFAP)免疫荧光染色法鉴定星形胶质细胞;实验分为对照组,模型组和2.5、5、10μmol·L-1丹皮酚组,0.5 mg·L-1脂多糖(LPS)诱导炎症反应。采用ELISA法测定培养液中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平;采用Western blot检测细胞IκBα蛋白表达和磷酸化水平及胞核NF-κB(p65)蛋白表达水平。结果与对照组相比,模型组星形胶质细胞上清液中IL-1β、IL-6和TNF-α水平显著增加(P<0.01),胞浆IκBα蛋白表达受抑(P<0.01),IκBα蛋白磷酸化和胞核NF-κB(p65)蛋白表达水平上调(P<0.01);5、10μmol·L-1丹皮酚能减少LPS活化的星形胶质细胞上清液中IL-1β、IL-6和TNF-α水平(P<0.05或P<0.01),增加胞浆IκBα蛋白表达(P<0.05或P<0.01),抑制LPS上调的IκBα蛋白磷酸化和胞核NF-κB(p65)蛋白表达水平(P<0.05或P<0.01)。结论丹皮酚能抑制LPS诱导的星形胶质细胞炎性因子IL-1β、IL-6和TNF-α的分泌,IκBα/NF-κB信号通路可能参与了丹皮酚对星形胶质细胞炎症反应的抑制作用。
METHODS Cultured astrocytes identification was evaluated by immunocytochemically stained with glial fibrillary acidic protein (GFAP). The cells were divided into control group, model group, and the 2.5, 5 and 10 μmol·L^-1 paeonol groups. LPS (0.5 mg·L^-1) was used to induce the inflammation. Proinflammatory cytokines IL-1β, IL-6 and TNF- α were determined by ELISA. Western blot was performed to observe the protein expression and phosphorylation levels of IκBα, and the protein expression of nuclear NF-κB (p65) in cultured astrocytes. RESULTS The levels of IL-1β, IL-6 and TNF- α in cell culture supernatant were significantly increased in the model group (P 〈 0.01 ), and the phosphorylation levels of IκBα and the protein expression of NF- κB (p65) were up regulated (P 〈 0.01 ), and the protein expression of IκBα in cultured astrocytes was down regulated (P 〈 0.01). The levels of IL-1/3, IL-6 and TNF-α in cell culture supernatant were significantly decreased ( P 〈 0.05 or P 〈 0.01) , LPS induced overexpression of NF - κB (p65) and phosphorylation levels of IκBα were inhibited (P 〈 0.05 or P 〈 0.01 ), and the protein expression of IκBα was increased in the 5, 10 μmol·L^-1 paeonol groups (P 〈 0.05 or P 〈 0.01). CONCLUSION Paeonol can inhibit the release of inflammatory cytokines induced by LPS in cultured astrocytes through the suppression of IκBα/NF- κB signal transduction pathway activity.
出处
《中国新药与临床杂志》
CAS
CSCD
北大核心
2014年第8期573-577,共5页
Chinese Journal of New Drugs and Clinical Remedies
基金
国家自然科学基金资助项目(81274134)
安徽省自然科学基金(1408085MH146)
安徽省高校省级自然科学研究项目(KJ2012A243
KJ2013Z275
KJ2014A202)
合肥师范学院生物化学与分子生物学扶持重点学科资助项目
关键词
牡丹皮
丹皮酚
脂多糖类
星形细胞瘤
炎症
细胞因子类
NF-κB
Cortex Moutan Redicis
paeonol
lipopolysaccharides
astrocytoma
inflammation
cyto- kines
NF- kappa B